Structural and chemical characterization of the S layer of a Pseudomonas-like bacterium.
Sections and freeze-fractured preparations showed an S layer on the surface of Pseudomonas-like strain EU2. Polyacrylamide gel electrophoresis of cell envelopes extracted with 1% sodium dodecyl sulfate (SDS) at room temperature showed three proteins (45K, 55K, and 110K). The 55K protein was identified as the S-layer protein. Incubation in 1.5 M guanidine hydrochloride removed the S layer from cell envelopes and dissociated the structure into subunits. The soluble 55K protein reassembled into planar sheets upon removal of the guanidine hydrochloride by dialysis. Electron microscopy and image processing indicated that these sheets had p4 symmetry in projection with a lattice constant of 13.2 +/- 0.1 nm (corresponding to 9.3 nm between adjacent fourfold axes). In some instances these reassemblies appeared to form small three-dimensional crystals which gave particularly clear views of the structure in projection because of the superimposition of information from a number of layers. A model is proposed with molecules having rounded lobes connected by a narrower linker region and joining at the lobes to form the fourfold axes of the array. The pattern superficially resembles those of other bacterial S layers, such as those of Aeromonas salmonicida, Aeromonas hydrophila, and Azotobacter vinelandii. Extraction of cell envelopes with 1% SDS at 50 degrees C released the 110K protein from the envelopes and removed an amorphous backing layer from the S layer. The 45K protein displayed heat-modifiable migration in SDS-polyacrylamide gel electrophoresis and was insoluble in SDS at 50 degrees C or in high concentrations of guanidine hydrochloride, suggesting that it was associated with the peptidoglycan.
Austin
J W
JW
Department of Microbiology and Immunology, University of Western Ontario, London, Canada.
Stewart
M
M
Murray
R G
RG
D001419:Bacteria / Q000648:ultrastructure*; D002462:Cell Membrane / Q000648:ultrastructure*; D005613:Freeze Etching; D005614:Freeze Fracturing; D008565:Membrane Proteins / Q000032:analysis / Q000302:isolation & purification; D008854:Microscopy, Electron; D008961:Models, Structural; D008970:Molecular Weight; D011549:Pseudomonas / Q000648:ultrastructure*
D008565:Membrane Proteins